Publications by authors named "Mohammad Fazilati"

36 Publications

Green Synthesis of Magnetic Nanoparticles Using Satureja hortensis Essential Oil toward Superior Antibacterial/Fungal and Anticancer Performance.

Biomed Res Int 2021 19;2021:8822645. Epub 2021 Jan 19.

Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan.

The biological synthesis of nanoparticles, due to their environmental and biomedical properties, has been of particular interest to scientists and physicians. Here, iron nanoparticles (FeNPs) were synthesized using essential oil. Then, the chemical, functional, and morphological properties of these nanoparticles were characterized by typical experiments such as Uv-Vis, FTIR, XRD, FE-SEM, PSA, zeta potential, EDX, and EDX mapping. The results indicated Fe nanoparticles' formation with a cubic morphological structure and a particle size in the range of 9.3-27 nm. The antimicrobial effects of these nanoparticles were further evaluated using disc diffusion, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungal concentration (MFC) against two gram-positive bacterial strains ( and ), two gram-negative bacterial strains ( and ), and one fungus species . The results showed that green-synthesized Fe nanoparticles possessed higher antimicrobial properties than Satureja hortensis essential oil against selected pathogenic microorganisms, especially Gram-negative bacteria. Finally, the anticancer effect of these Fe nanoparticles was investigated on human cancer cells, K-562, and MCF-7, by the MTT assay. The results showed the anticancer effect of these nanoparticles against selected cell lines.
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http://dx.doi.org/10.1155/2021/8822645DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7840253PMC
January 2021

High-level Expression and One-step Purification of Chimeric Antigen Containing HTLV-I-II Diagnostic Epitopes in Escherichia coli.

Iran J Allergy Asthma Immunol 2020 Apr 16;19(2):149-158. Epub 2020 Apr 16.

Department of Biochemistry, Payame Noor University, Tehran, Iran.

Purification and preparation of three diagnostic antigens used for the detection of human T-lymphotropic virus (HTLV)-I/-II infection in E.coli are different parts of a multi-step method. In this study, our aim was to design a chimeric protein for the simultaneous detection of HTLV-I and HTLV-II antibodies. Immunodominant B cell linear epitopes of envelope and capsid proteins of HTLV-I/-II were selected and linked together; using a suitable amino acid linker and a chimeric antigen (CA). The codon-optimized synthetic DNA encoding the CA was subcloned into the pGS21aexpression vector and CA expressed as His-GST fused protein in E. coli BL21 (DE3) cells. Then the recombinant CA was purified, using the Ni-NTA (Nickle Nitrilotriacetic acid) affinity chromatography under native conditions. The Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and densitometric scanning results showed that CA accounted for 15% of the total cellular proteins and approximately 50% of the expressed histidine-glutathione s-transferase-chimeric antigen (His-GST-CA) proteins were soluble. The CA was successfully purified in one step with a purity of greater than 90%, which is suitable for antigenicity evaluations. Enzyme-linked immunosorbent assay (ELISA) results showed that the GST fused CA reacted in a concentration-dependent manner with HTLV-I/-II infected sera and was able to distinguish normal serum from HTLV-I/-II infected one with a proper sensitivity. With further validation, CA, as described in the present study could be introduced as a novel reliable, cost-effective and easy alternative for the three separate HTLV-I/-II diagnostic peptide antigens, which is prepared as a fusion with GST.
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http://dx.doi.org/10.18502/ijaai.v19i2.2765DOI Listing
April 2020

Association between serum cell adhesion molecules with hs-CRP, uric acid and VEGF genetic polymorphisms in subjects with metabolic syndrome.

Mol Biol Rep 2020 Feb 23;47(2):867-875. Epub 2019 Dec 23.

Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, 99199-91766, Iran.

Metabolic syndrome (MetS) is associated with a pro-inflammatory state and endothelial dysfunction that places subjects with MetS at a higher risk of atherosclerosis. Inflammatory biomarkers are raised in patients at risk of developing cardiovascular diseases. In the current study, we aimed to examine the possible association between MetS and serum soluble adhesion molecules, hs-CRP, uric acid, and the genetic variations related to vascular endothelial growth factor (VEGF) gene. In this cross-sectional study, participants were enrolled from the Mashhad stroke and heart atherosclerotic disorders (MASHAD) study. The International Diabetes Federation criteria were used to define the MetS. Cell adhesion molecules (CAM) and serum hs-CRP were measured by ELISA and PEG-enhanced immunoturbidimetry method, respectively. We used a logistic regression analysis to determine independent associations of CAMs with the VEGF polymorphisms and MetS. Two hundred and 59 participants with and without MetS were enrolled. Participants with MetS and DM had a significantly higher serum E-selectin level (p < 0.05). Participants with a high serum E-selectin level had higher levels of hs-CRP, FBG, TG, uric acid, BMI and lower levels of serum HDL-C (p < 0.05). Interestingly, individuals with MetS with a genetic variant of the VEGF gene (rs6921438) had higher level of serum ICAM-1 (p = 0.04). There were significant associations between serum E-selectin concentrations and the presence of MetS, and its risk factors. Moreover, we demonstrated that MetS subjects with the rs6921438 genetic variant had a higher serum level of ICAM-1 (p < 0.05).
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http://dx.doi.org/10.1007/s11033-019-05081-2DOI Listing
February 2020

Variation of ursolic acid content in flowers of ten Iranian pomegranate ( L.) cultivars.

BMC Chem 2019 Dec 3;13(1):80. Epub 2019 Jul 3.

4Department of Chemistry, Payame Noor University (PNU), Tehran, Iran.

Background: Ursolic acid (UA) is an important bioactive component in many traditional medicinal plants including pomegranate ( L.) flower.

Methods: This study presents the HPLC analysis of UA contents of ten cultivars of pomegranate flower grown in Iran. The UA contents of fallen flowers of pomegranate were given in each cultivar.

Results: Remarkable quantities of UA were found in ten cultivars of Iranian pomegranate flower evaluated (21.736 to 15.119 mg/g). Lower quantities of UA were determined in pomegranate fallen flowers (16.763 to 5.754 mg/g).

Conclusion: UA values obtained from Iranian cultivars of pomegranate flowers are very significant when compared with other sources of UA. All of the analyzes suggested that the Iranian pomegranate flowers (including flowers on branches and fallen flowers) might be an excellent UA rich source.
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http://dx.doi.org/10.1186/s13065-019-0598-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6661776PMC
December 2019

Evaluation of antibacterial property of hydroxyapatite and zirconium oxide-modificated magnetic nanoparticles against and .

IET Nanobiotechnol 2019 Jun;13(4):449-455

Biology Department, University of Isfahan, Isfahan, Iran, Iran.

In the first section of this research, superparamagnetic nanoparticles (NPs) (FeO) modified with hydroxyapatite (HAP) and zirconium oxide (ZrO) and thereby FeO/HAP and FeO/ZrO NPs were synthesised through co-precipitation method. Then FeO/HAP and FeO/ZrO NPs characterised with various techniques such as X-ray photoelectron spectroscopy, X-ray diffraction, scanning electron microscopy, energy dispersive X-ray analysis, Brunauer-Emmett-Teller, Fourier transform infrared, and vibrating sample magnetometer. Observed results confirmed the successful synthesis of desired NPs. In the second section, the antibacterial activity of synthesised magnetic NPs (MNPs) was investigated. This investigation performed with multiple microbial cultivations on the two bacteria; () and (). Obtained results proved that although both MNPs have good antibacterial properties, however, FeO/HAP NP has greater antibacterial performance than the other. Based on minimum inhibitory concentration and minimum bactericidal concentration evaluations, bacteria are more sensitive to both NPs. These nanocomposites combine the advantages of MNP and antibacterial effects, with distinctive merits including easy preparation, high inactivation capacity, and easy isolation from sample solutions by the application of an external magnetic field.
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http://dx.doi.org/10.1049/iet-nbt.2018.5029DOI Listing
June 2019

ErbB4 receptor polymorphism 2368A>C and risk of breast cancer.

Breast 2018 Dec 9;42:157-163. Epub 2018 Oct 9.

Division of Cellular and Molecular Biology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran; Department of Cellular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran. Electronic address:

Purpose: A number of single nucleotide polymorphisms (SNPs) in EebB4 gene have been studied, which has clarified their impact on breast cancer in different populations. Nevertheless, the importance of rs13423759 in breast cancer has not been studied and its effect remained almost unclear. In this paper, we evaluated the frequency of rs13423759 different alleles in Iranian population and statistically analyzed their association with breast cancer risk.

Materials And Methods: Allele-specific Primer PCR (ASP-PCR) was recruited in this study to genotype rs13423759 position in 172 breast cancer and 148 healthy control subjects. The genotypes of control and cases were analyzed statistically to find the association between rs13423759 alleles and breast cancer incidence and its clinicopathological characteristics. In silico studies were performed in order to find the mechanistic viewpoint of rs13423759 alleles in breast cancer.

Results: rs13423759 allele C was shown to be significantly associated with breast cancer risk, HER2 positivity and increased risk of metastasis. Reciprocally, allele A was correlated with the lowered risk of breast cancer. The in silico studies showed that rs13423759 allele C is capable to strengthen the interaction between miR-548as, an oncomiRNA, and ErbB4 mRNA, leading to its lowered concentration in the cells.

Conclusion: rs13423759 allele C is significantly associated with the enhanced risk of breast cancer, elevated metastasis and HER2 positivity.
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http://dx.doi.org/10.1016/j.breast.2018.10.002DOI Listing
December 2018

Amyloid Nano-biofibrils as a New Nano-Scaffold for Lipase Immobilization.

Protein Pept Lett 2018 ;25(9):862-870

Department of Biology, Payame Noor University, Tehran, Iran.

Background: Amyloids could be created under destabilizing conditions from various proteins. Having high chemical reactive groups makes the amyloid fibers suitable for enzyme stabilization. Imobilization of lipase as one of the stable classes of high catalytic power enzymes could be very valuable.

Objective: In the present study, the lipase from Pseudomonas cepacia was immobilized on BSA amyloid nano-biofibrils and the kinetic parameters were compared with those of its free counterpart. The possibility of using this nano-material as a new nano-scaffold for lipase immobilization was investigated.

Method: Response surface methodology was used in this study to produce the maximum amounts of amyloid fibrils using Design Expert 7 software. Transmission electron microscopy was employed to confirm the presence of amyloid fibers. The stabilization process was performed by glutaraldehyde mediated covalent cross-links between the enzyme and amyloid fibers. Kinetic parameters including activity, specific activity, optimal pH and temperature and thermal stability of immobilized enzyme were compared with the free counterpart.

Results: The optimum conditions for fibrillogenesis were obtained at 4.36 mg.ml-1 of protein after 72 hours of mild agitation in a mixed citrate-phosphate buffer at the pH of 4.5 and the temperature of 80 ºC. The kinetic parameters of the immobilized lipase were improved in terms of activity, specific activity, Km and Vmax, optimal pH and temperature and thermal stability at 40 ºC. Amyloid fibrils with a diameter of less than 100 nm, as a new nano-scaffold, increased both the stability of lipase and other kinetic properties of the enzyme.

Conclusion: Amyloid fibrils as a new chemically-rich nano-scaffold could be an appropriate matrix for lipase immobilization.
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http://dx.doi.org/10.2174/0929866525666180911155312DOI Listing
January 2019

The effect of , , and polymorphism in patients under warfarin therapy in city of Kermanshah.

Res Pharm Sci 2018 Aug;13(4):377-384

Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, I.R. Iran.

Polymorphism in the genes encoding enzyme and reductase significantly influence warfarin dose requirement since patients with , and mutant alleles require lower warfarin maintenance doses. Studies have reported the ethnic variations in the frequency of these genes within the various populations in Iran and other parts of the world. However, no such study has been done yet on Kurdish population in Kermanshah. From Kurdish population of Kermanshah province in Iran, a total of 110 patients who had heart surgery and taking warfarin, were genotyped for polymorphisms of , , and . Polymorphism genotyping was performed by sequencing as well as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using restriction enzymes of and , respectively. The frequencies of -1639 GG, GA, and AA genotypes were 42%, 36%, and 22%, respectively and for 1*/1*, 1*/2*, 2*/2*, 1*/3*, 3*/3*, 2*/3* were 71%, 17%, 5.4%, 1.8%, 4.5%, and 0%, respectively. The frequency of -1639A allele was 42.3% and the frequencies of and *3 alleles were 14% and 5.4%, respectively. It was indicated that low warfarin dose requirements are strongly associated with the presence of and variant alleles. Our results confirmed the supply to understand the distribution of genomic biomarkers related to the drugs metabolism for future planning health programs.
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http://dx.doi.org/10.4103/1735-5362.235165DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6040167PMC
August 2018

An investigation of the association between the level of prolactin in serum and type II diabetes.

Diabetes Metab Syndr 2019 Sep - Oct;13(5):3035-3041. Epub 2018 Jul 17.

Department of Clinical Biochemistry, Faculty of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran. Electronic address:

As a hormone secreted from the pituitary gland, prolactin (PRL) plays an important role in increasing beta cell proliferation, stimulating the secretion of insulin, preventing the activities of caspases on pathways that cause apoptosis in the Langerhans' islands, and moderating the immune system in regulating the whole body's sensitivity to insulin. Therefore, PRL level changes in type II diabetes and it can be concluded that PRL can play an important role in metabolic disorders of glucose. The present study is carried out in order to investigate the association between serum levels of PRL and type II DM. Blood samples were taken from 64 females affected by type II diabetes and 70 healthy ones, whose PRL level was measured using electrochemiluminescence (ECL) technique. It was a case-control study, and based on the definition dedicated to each group, subjects were assigned to two groups. The patient group included the subjects with type II diabetes while the control group included healthy samples. Data were analyzed using SPSS software (Mann-Whitney test, t-test, and spearman's rho correlation test). According to the results, PRL concentration in the serum of people affected by type II diabetes (5.32 ± 0.36) was significantly (P˂0.05) lower than that of control group (18.38 ± 2.3). The results also showed that in type II diabetes, the level of PRL changes so that the concentration of PRL in the serum of the patients was lower than that of healthy ones. Therefore, PRL concentration in the blood can be related to diabetes.
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http://dx.doi.org/10.1016/j.dsx.2018.07.007DOI Listing
February 2020

Association of rs6921438 A

Gene 2018 08 4;667:70-75. Epub 2018 May 4.

Department of Modern Sciences and Technologies, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Metabolic Syndrome Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Background: Genome-wide association studies (GWAS) have identified common variants at the Vascular-Endothelial-Growth-Factor (VEGF) gene locus, which appear to be associated with plasma VEGF concentrations. These factors are among the major risk factors for cardiovascular disease and metabolic syndrome (MetS). We have investigated the association between serum VEGF concentrations and a VEGF genetic variant (rs6921438 A
Methods: In total of 852 individuals, genotyping was performed using polymerase-chain-reaction and restriction-fragment-length-polymorphisms. Serum VEGF level was determined in 122 subjects using the EV 3513 cytokine biochip array. Anthropometric and biochemical characteristics, including fasting blood glucose and lipid profile evaluated by univariate and multivariate analyses. The association of the polymorphism with serum VEGF level and its interaction with dietary intake in association with the essential determinants of cardiovascular risk factors were assessed.

Results: As would be expected, patients with MetS had markedly higher body mass index, waist-circumference, serum total cholesterol, triglyceride, hs-CRP and blood pressure, and lower concentrations of HDL-C, compared to non-MetS individuals (P < 0.05). The association between the rs6921438 single nucleotide polymorphism (SNP) and the presence of MetS and individual features of MetS were not statistically significant. Interestingly we observed a significant association between high serum VEGF levels with the GG and GA genotypes in the individuals with MetS, compared to the wild-type genotype, which was also associated with dietary fat intake.

Conclusion: Our findings show an association between a VEGF gene polymorphism with serum VEGF concentrations and dietary fat intake, but there was no association with the presence of MetS.
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http://dx.doi.org/10.1016/j.gene.2018.05.017DOI Listing
August 2018

Concentration of lead and mercury in collected vegetables and herbs from Markazi province, Iran: a non-carcinogenic risk assessment.

Food Chem Toxicol 2018 Mar 31;113:204-210. Epub 2018 Jan 31.

Department of Food Science, Faculty of Food Engineering, University of Campinas (UNICAMP), Campinas, Sao Paulo, Brazil. Electronic address:

The current study was undertaken to determine the concentration of Hg and Pb in ten types of collected green leafy vegetables and herbs from different agricultural sites of Markazi province, Iran as well as the gathered water and soil around them using inductively coupled plasma-optical emission spectroscopy (ICP-OES). Also, the potential health risk assessment by using target hazard quotient (THQ) and hazard index (HI) parameters was estimated. Based on the accumulation order, Artemisia dracunculus L with 56.147 ± 17.30 μg/kg and Spinacia oleracea L with 1733.62 ± 2264.7 μg/kg can uptake and accumulate more concentration of Pb and Hg in their tissues, respectively. Regarding gathered soil around vegetables, the concentration of Hg and Pb were measured as 52.056 ± 16.25 μg/kg and 4993.83 ± 1287.8 μg/kg, respectively. The transfer factor (TF) demonstrated that vegetables and herbs could absorb a high amount of Hg from the soil while these plants uptake less concentration of Pb thought their green leaves. The non-carcinogenic risk assessment showed the minimum, and maximum THQ was related to 15-24 and 35-44 age groups in the urban and rural consumers. Also, HI in the urban and rural areas was calculated as 8.492 and 9.012, respectively. Since HI > 1, exposure of the urban and rural areas of Markazi province to non-carcinogenic risk by consuming the green leafy vegetables and herbs is a source of concern.
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http://dx.doi.org/10.1016/j.fct.2018.01.048DOI Listing
March 2018

The Effects of Flaxseed Oil Omega-3 Fatty Acids Supplementation on Metabolic Status of Patients with Polycystic Ovary Syndrome: A Randomized, Double-Blind, Placebo-Controlled Trial.

Exp Clin Endocrinol Diabetes 2018 Apr 8;126(4):222-228. Epub 2017 Nov 8.

Research Center for Biochemistry and Nutrition in Metabolic Diseases, Kashan University of Medical Sciences, Kashan, I.R. Iran.

Objective: This study was conducted to evaluate the effects of flaxseed oil omega-3 fatty acids supplementation on metabolic status of patients with polycystic ovary syndrome (PCOS).

Methods: This randomized double-blind, placebo-controlled trial was conducted on 60 women with PCOS according to the Rotterdam criteria aged 18-40 years old. Participants were randomly assigned into two groups to receive either 1,000 mg flaxseed oil omega-3 fatty acids (n=30) or placebo (n=30) twice a day for 12 weeks. Metabolic, endocrine, inflammatory factors were quantified at baseline and after the 12-week intervention.

Results: After the 12-week intervention, compared to the placebo, flaxseed oil omega-3 supplementation significantly decreased insulin values (-2.6±7.7 vs.+1.3±3.9 µIU/mL, P=0.01), homeostasis model of assessment-estimated insulin resistance (-0.7±1.7 vs.+0.3±0.9, P=0.01), mF-G scores (-1.2±1.7 vs. -0.1±0.4, P=0.001), and increased quantitative insulin sensitivity check index (+0.01±0.02 vs. -0.01±0.02, P=0.01). In addition, supplementation with flaxseed oil omega-3 resulted in significant decreases in serum triglycerides (-5.1±20.9 vs.+9.7±26.1 mg/dL, P=0.01), VLDL-cholesterol (-1.0±4.2 vs.+1.9±5.2 mg/dL, P=0.01) and high-sensitivity C-reactive protein (hs-CRP) (-1.6±3.1 vs.+0.2±1.5 mg/L, P=0.004) compared to the placebo. We did not see any significant effect of flaxseed oil omega-3 supplementation on hormonal and other lipid profiles, and plasma nitric oxide levels.

Conclusions: Overall, flaxseed oil omega-3 supplementation for 12 weeks in women with PCOS had beneficial effects on insulin metabolism, mF-G scores, serum triglycerides, VLDL-cholesterol and hs-CRP levels, but did not affect hormonal and other lipid profiles, and plasma nitric oxide levels.
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http://dx.doi.org/10.1055/s-0043-119751DOI Listing
April 2018

Inhibitory action of dicyclomine on lipase activity, kinetics and molecular study.

Int J Biol Macromol 2018 Feb 18;107(Pt B):2422-2428. Epub 2017 Oct 18.

Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Lipase is one of the most important groups of enzymes for industry and medicine. It breaks down triacylglycerol to glycerol and fatty acids. Some bacteria use lipase to degrade the extracellular matrix of the host cells to penetrate into the tissues. Dicyclomine is a muscarinic antagonist receptor that relieves the smooth muscle spasm of the gastrointestinal tract and affects the cardiovascular system. In this research, the effect of a dicyclomine on the lipase activity of Pseudomonas aeruginosa was studied. Hanes-Woolf plot showed that the drug inhibited the enzyme by competitive inhibition. The IC value (60uM) and Ki (30uM) of the drug revealed that the drug bound to enzyme with high affinity. Determination of enzyme activity in various temperature showed that the maximum activity of lipase was at 60°C both in the presence and absence of the drug. Arrhenius plot determined that the activation energy of the enzyme reaction was increased in the presence of the drug. The model of binding demonstrated that the drug entered a pocket containing 10 amino acids and interacted by hydrogen bond and hydrophobic interaction and the conformational change of the enzyme after binding of the drug was confirmed by fluorescence measurement.
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http://dx.doi.org/10.1016/j.ijbiomac.2017.10.123DOI Listing
February 2018

The association between rs1972820 and the risk of breast cancer in Isfahan population.

J Cancer Res Ther 2017 Jan-Mar;13(1):26-32

Department of Biochemistry, Payame Noor University of Isfahan, Isfahan, Iran.

Context: A number of single nucleotide polymorphisms (SNPs) in ERBB4 gene have been linked to increase the risk of breast cancer. However, no study has been dedicated to analyze the significance of microRNA-related SNP rs1972820, located in ERBB4 3'-untranslated region (UTR), in breast tumors.

Aims: Here, we investigated the frequency and association between rs1972820 and breast cancer.

Subjects And Methods: The rs1972820 genotypes in 182 samples were collected from 96 healthy people, and 86 breast cancer patients were determined using tetra-primer amplification refractory mutation system-polymerase chain reaction. The frequency of genotypes was analyzed to find the association between rs1972820 and breast cancer risk.

Statistical Analysis Used: Conditional logistic regression, odds ratios (ORs), the associated 95% confidence intervals (CIs), and Armitage's test were used in this study.

Results: In silico analysis suggested that rs1972820 located in the 3'UTR of ERBB4 gene affects the binding affinity of miR-3144-3p a potential oncomiRNA. Statistical analysis showed a significant association between SNP rs1972820 G allele and reduced breast cancer risk, odds ratio = 0.443 (95% CI: 0.196-0.998).

Conclusions: rs1972820 SNP allele is significantly associated with the reduced risk of breast cancer and could be considered as a potential marker for breast cancer predisposition in population of Isfahan.
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http://dx.doi.org/10.4103/0973-1482.183202DOI Listing
February 2018

Transcription factor 7-like 2 polymorphism and context-specific risk of metabolic syndrome, type 2 diabetes, and dyslipidemia.

J Res Med Sci 2017 15;22:40. Epub 2017 Mar 15.

Department of Biochemistry, Isfahan Payame Noor University, Isfahan, Iran.

Background: The transcription factor 7-like 2 gene (TCF7L2) is an element of the Wnt signaling pathway. There is lack of evidence if TCF7L2 has a functional role in lipid metabolism and regulation of the components constitutes the metabolic syndrome (MetSyn). The aims of this study were to evaluate whether the risk allele of TCF7L2 gene polymorphism is associated with dyslipidemia and MetSyn.

Materials And Methods: The MetSyn subjects were participated only based on the National Cholesterol Education Program - Third Adult Treatment Panel criteria. In this case-control study, the DNA from MetSyn patients without ( = 90) and with type 2 diabetes (T2D) ( = 94) were genotyped.

Results: The results show that the genotype-phenotype for CC, CT/TT of TCF7L2 gene polymorphism correlated with body mass index and waist circumference in MetSyn and MetSyn + T2D subjects ( = -0.949 and = -0.963, respectively). The subjects that only possess MetSyn but are not diabetics show the 2 h postprandial glucose and fasting blood glucose, glycated hemoglobin significantly lower ( < 0.05) than those subjects have both abnormality. The level of triglyceride in CT/TT carriers in MetSyn was higher than CC carriers ( = 0.025). A comparison with the controls subjects, the frequencies of the T allele in the groups of MetSyn (46.66%) and MetSyn + T2D (47.34%) show significantly different ( < 0.05). The odds ratios for T allele in (MetSyn)/(normal), (MetSyn + T2D)/(normal), and in (MetSyn + T2D)/(MetSyn) were 3.59 (95% confidence interval [CI], 1.33-9.67, = 0.0093), 3.76 (95% CI, 1.40-10.07, = 0.0068), and 1.08 (95% CI: 0.55- 2.11, = 0.834), respectively.

Conclusion: The results revealed the important insights essential for the role of TCF7L2 that the T allele of TCF7L2 plays a significant role in the susceptibility to dyslipidemia, MetSyn, and T2D.
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http://dx.doi.org/10.4103/1735-1995.202141DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5393097PMC
March 2017

Novel carboxymethyl cellulose-polyvinyl alcohol blend films stabilized by Pickering emulsion incorporation method.

Carbohydr Polym 2017 Jul 9;167:79-89. Epub 2017 Mar 9.

Young Researchers and Elite Club, Hamedan Branch, Islamic Azad University, Hamedan, Iran. Electronic address:

The aim of this study was to investigate the possibility of increasing the antimicrobial and antioxidant properties of biodegradable active films stabilized via Pickering emulsions. The blend films were prepared from carboxymethyl cellulose (CMC) and polyvinyl alcohol (PVA), emulsified with oleic acid (OL) and incorporated with rosemary essential oil (REO). Formation of Pickering emulsion was confirmed by scanning electron microscopy (SEM), optical microscopy, mean droplet size and emulsion stability. Morphological, optical, physical, mechanical, thermal, antifungal and antioxidant properties of the films incorporated with different concentrations of REO (0.5, 1.5 and 3%) were determined. The results showed an increase in UV absorbance and elongation at break but, a decrease in tensile strength and thermal stability of the films. Interestingly, films containing REO exhibited considerable antioxidant and antimicrobial properties. In vitro microbial tests exhibited 100% fungal inhibition against Penicillium digitatum in the films containing 3% REO. In addition, no fungal growth were observed after 60days of storage at 25°C in bread slices were stored with active films incorporated with 3% REO, could attributed to the slow and regular release of REO caused by Pickering emulsions. The results of this study suggest that Pickering emulsion is a very promising method, which significantly affects antioxidant and antimicrobial activities of the films.
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http://dx.doi.org/10.1016/j.carbpol.2017.03.017DOI Listing
July 2017

A Novel Mutation in SLC7A9 Gene in Cystinuria.

Iran J Kidney Dis 2017 Mar;11(2):138-141

Pediatric Inherited Diseases Research Center, Research Institute for Primordial Prevention of Non-communicable Disease and Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Introduction: Cystinuria is an inherited disorder affecting luminal transport of cystine and dibasic amino acids. Because of the poor solubility of cystine in urine, stone formation in the kidney occurs frequently. Cystinuria is associated with mutations in the SLC3A1 and SLC7A9 genes. Despite the population-specific distribution of mutations in the SLC7A9 genes, there are few genetic data reported for cystinuric patients from the Middle East.

Materials And Methods: Exon 4 of the SLC7A9 gene was sequenced in 21 patients with cystinuria, using the polymerase chain reaction and sequencing methods.

Results: A new variation in exon 4 of the SLC7A9 gene was identified, which was insertion of 1 adenine nucleotide between 2 cytosine nucleotides in position c.213-214insA.

Conclusions: It seems to be important since it causes frame shift and it may be an important cause to make disease.
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March 2017

Aqueous extract of Launaea acanthodes induces glutamate uptake and GABA release in astrocyte cell culture via a ROS scavenging mediated process.

J Chem Neuroanat 2017 Jul 24;82:1-4. Epub 2017 Feb 24.

Department of Biology, Payame Noor University, Isfahan, Iran.

Launaea acanthodes is extensively used in the semiarid region of Iran for treatment of seizure. However, the underlying mechanism has not been studied well. In our previous study we showed that Launaea acanthodes extract could effectively stimulate GABA release from PC12 cell culture. The critical role of astrocytes in epileptic brain in regulation of neurotransmitter balance in central nervous system encouraged us to investigate the effect of Launaea acanthodes extract on GABA and glutamate release from astrocytes. Our results indicated that LA extract could stimulates both glutamate uptake and GABA release by astrocytes. The results confirmed this fact that GABA release by astrocytes in response to LA treatment is a glutamate uptake-dependent process. We showed that stimulation of GABA release by Launaea acanthodes is a gene expression based process which depends on glutamate uptake. We propose that glutamate uptake via glutamate transporter 3 could activate expression of glutamate decarboxylase which in turn transforms uptaken glutamate into GABA.
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http://dx.doi.org/10.1016/j.jchemneu.2017.02.008DOI Listing
July 2017

ESR1 single nucleotide polymorphism rs1062577 (c.*3804T>A) alters the susceptibility of breast cancer risk in Iranian population.

Gene 2017 May 23;611:9-14. Epub 2017 Feb 23.

Biochemistry Division, Department of Biology, Faculty of Science, Payame Noor University of Taft, Yazd, Iran.

Objectives: Albeit single nucleotide polymorphisms related to ESR1 gene have been studied, only a number of them have been reported to be associated with breast cancer risk. rs1062577 is one of the most recent microRNA-related ESR1 SNPs; however, no study has been conducted to investigate the significance this polymorphism in Iranian population. In this study, we aimed to investigate the frequency and also the association between rs1062577 and breast cancer.

Materials And Methods: rs1062577 position was genotyped by Tetra-primer ARMS-PCR in totally 182 blood specimens obtained from breast cancer patients (n=86), and healthy blood donors (n=96). The distribution of different genotypes was statistically analyzed in terms of the potential association between rs1062577 different alleles, breast cancer risk and clinicopathological criteria of breast cancer patients.

Results: The statistical analyses confidently indicated that rs1062577 A allele is associated with the increased breast cancer risk in both univariate and multivariate regression models (Odds Ratio=8.403 and 32.602 respectively). rs1062577 T allele was statistically associated with stage I of breast cancer patients (p-value=0.025). In silico studies implied that rs1062577 A allele can alter the binding capacity of ESR1 mRNA and miRNAs via either breakage or formation of hydrogen bonds.

Conclusion: rs1062577 A allele is significantly and dramatically associated with the elevated risk and greater stages of breast cancer.
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http://dx.doi.org/10.1016/j.gene.2017.02.016DOI Listing
May 2017

Association of a Vascular Endothelial Growth Factor genetic variant with Serum VEGF level in subjects with Metabolic Syndrome.

Gene 2017 Jan 27;598:27-31. Epub 2016 Oct 27.

Molecular Medicine Group, Department of Modern Sciences and Technologies, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Biochemistry of Nutrition Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Background: The metabolic syndrome (MetS) is a clustering of metabolic disorders that is associated with an increased risk of developing cardiovascular-disease, diabetes, and related diseases. Against this background, Vascular Endothelial Growth Factor (VEGF) plays an essential role in angiogenesis, vascular permeability, and hematopoiesis and its increased level is reported to be associated with increasing the risk of developing cardiovascular-disease, stroke and diabetes. Therefore the aim of present study was to explore the association of serum VEGF level and its associated genetic-polymorphism, rs10738760 (A>G) at 9p24.2, in 850 subjects with/without MetS.

Methods: MetS was defined according to the International-Diabetes-Federation criteria. Genotyping was carried out using Polymerase chain reaction-amplification refractory mutation system. Anthropometric/biochemical parameters, including FBG, Triglyceride, HDL, TC, etc., were determined followed by univariate and multivariate analyses.

Results: MetS patients had significantly higher levels of BMI, waist-circumference, cholesterol, triglyceride, Hs-CRP and SBP/DBP, while the HDL-C levels was lower in patients group, compared to control group (P<0.05). Moreover, our analysis showed that MetS patients with GA or AA genotypes had a significantly (P=0.03) higher serum level of VEGF.

Conclusions: we demonstrate an association between a VEGF genetic variant with MetS, suggesting its role as a risk stratification factor for MetS.
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http://dx.doi.org/10.1016/j.gene.2016.10.034DOI Listing
January 2017

Leptin and leptin-receptor polymorphisms in fertile and infertile men.

Syst Biol Reprod Med 2017 Feb 16;63(1):7-14. Epub 2016 Dec 16.

f Urology and Nephrology Research Center , Hamadan University of Medical Sciences , Hamadan , Iran.

The association of leptin (LEP) -2548G/A and/or leptin receptor (LEPR) Gln223Arg polymorphisms with male infertility and plasma FSH, LH, and testosterone (T) levels was examined. The genotypes and allele frequency distributions of LEP -2548G/A and LEPR Gln223Arg polymorphisms were investigated in 150 fertile and 150 infertile men by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Also, plasma levels of FSH, LH, and T were measured using commercial ELISA kits. Frequencies of AA, AG and GG genotypes of LEP-2548G/A polymorphism were statistically different in fertile and infertile men (p=0.012). The AG genotype showed a protective effect which could decrease risk of male infertility about 3 fold (p = 0.004). We did not observe any differences in frequencies of LEPR Gln223Arg alleles and genotypes between groups (p > 0.05). Sperm counts from infertile men with the AG and GG genotypes of the LEP polymorphism were significantly higher than AA genotype (p<0.05). Moreover, infertile men who carried the RR genotype of LEPR showed a statistically higher percentage of sperm with progressive motility than individuals with other genotypes (p = 0.004). There was no correlation between different combinations of LEP and LEPR genotypes and LH, FSH, and T levels (p > 0.05). Our study suggests that the LEP -2548G/A polymorphism may play a role in male fertility and the AG genotype may have a protective effect through increasing sperm counts. The distribution of genotypes of LEP -2548G/A polymorphism are different in fertile and infertile males and may be a useful tool in evaluation of male infertility.

Abbreviations: LEP: leptin; LEPR: leptin receptor; T: testosterone; FSH: follicle-stimulating hormone; LH: luteinizing hormone.
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http://dx.doi.org/10.1080/19396368.2016.1258741DOI Listing
February 2017

Molecular detection of aminoglycoside-modifying enzyme genes in Acinetobacter baumannii clinical isolates.

Acta Microbiol Immunol Hung 2017 Jun 16;64(2):143-150. Epub 2016 Dec 16.

5 Department of Biology, Payame Noor University , Isfahan, Iran.

Acinetobacter baumannii is a major opportunistic pathogen in healthcare settings worldwide. In Iran, there are only few reports on the prevalence of aminoglycoside resistance genes among A. baumannii isolates. The aim of this study was to investigate the existence of aminoglycoside-modifying enzyme (AME) genes from A. baumannii strains collected at a university teaching hospital in Iran. One hundred A. baumannii strains were collected between 2014 and 2015 from hospitalized patients at Loghman Hakim Hospital, Tehran, Iran. Antimicrobial susceptibility was determined by disk diffusion method according to the Clinical and Laboratory Standards Institute recommendations. The DNA was extracted using a kit obtained from Bioneer Co. (Korea) and was used as a template for polymerase chain reaction. The most active antimicrobial agent against these strains was colistin. The rate of extended-spectrum cephalosporin resistance was 97%. The aadA1, aadB, aac(6')-Ib, and aac(3)-IIa genes were found in 85%, 77%, 72%, and 68% of A. baumannii isolates, respectively. This study showed a high prevalence rate of AME genes in A. baumannii. This prevalence rate has explained that further aminoglycoside resistance genes may have role in the resistance of clinical isolates of A. baumannii. Therefore, control and treatment of serious infections caused by this opportunistic pathogen should be given more consideration.
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http://dx.doi.org/10.1556/030.63.2016.022DOI Listing
June 2017

Polymorphisms in CD14 Gene May Modify Soluble CD14 Levels and Represent Risk Factors for Multiple Sclerosis.

Immunol Invest 2016 Nov 7:1-8. Epub 2016 Nov 7.

a Department of Neurology, School of Medicine , Isfahan University of Medical Sciences , Isfahan , Iran.

Background: Besides the central role of the adaptive immune system, a disturbance of innate immune system is also suggested to be involved in the pathogenesis of multiple sclerosis (MS). CD14, a receptor upregulated in activated microglia, is known to be an essential mediator of inflammation in innate immune responses. Therefore, in this study we aimed to assess possible roles of CD14-159 and -260 gene polymorphisms in MS susceptibility and the effects of those polymorphisms to its protein producing capacity in Iranian population.

Methods: In this case control study, CD14-159 and -260 polymorphisms were genotyped using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 200 MS patients and 200 healthy controls matched in age and gender. Serum levels of soluble CD14 (sCD14) was determined by enzyme-linked immunosorbent assay (ELISA).

Results: There were significant differences in genotype distribution of CD14-159 and -260 polymorphisms between patients and controls (P = 0.01, for-both). Mean serum level of sCD14 was significantly higher in MS patients than in control subjects (3340.30 ± 612.50 ng/ml vs 2353.73 ± 539.07 ng/ml; P < 0.01).

Conclusion: In summary, we conclude that CD14-159 and -260 polymorphisms are associated with the risk of MS in Iranian population and affects CD14 promoter activity, thereby regulating CD14 expression. Furthermore, our study provides preliminary evidence for the activation of innate immunity in the pathogenesis of MS. In addition, the findings of the present study suggest serum level of sCD14 as candidate biomarker of MS severity.
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http://dx.doi.org/10.1080/08820139.2016.1226897DOI Listing
November 2016

rs11895168 C allele and the increased risk of breast cancer in Isfahan population.

Breast 2016 Aug 1;28:89-94. Epub 2016 Jun 1.

Biochemistry Department, Payam-e Noor University of Isfahan, Isfahan, Iran.

Objectives: Some of the single nucleotide polymorphisms in EebB4 gene have been studied to date in order for finding their relevance to the risk of breast cancer. However, no study has been conducted to investigate the importance of rs11895168, a microRNA-related SNP located in ErbB4 3'UTR, in breast tumors. In this paper, we investigated the frequency and association between rs11895168 and breast cancer.

Materials And Methods: The rs11895168 was genotyped in 364 samples collected from 172 breast cancer patients and 192 healthy participants, using Tetra-primer ARMS PCR. The frequency of genotypes was analyzed to find the association between rs11895168 and breast cancer risk and also clinicopathological characteristics of patients.

Results: our in silico studies suggested that different alleles at rs11895168 affect the binding strength of miR-1276, a potential tumor suppressor. Statistical analysis showed a significant association between rs11895168 C allele-harboring genotypes and increased breast cancer risk.

Conclusion: rs11895168 C allele is strongly and significantly associated with the increased risk of breast cancer and positivity of ER/PR tumor cells.
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http://dx.doi.org/10.1016/j.breast.2016.05.007DOI Listing
August 2016

Gallic acid and exercise training improve motor function, nerve conduction velocity but not pain sense reflex after experimental sciatic nerve crush in male rats.

Avicenna J Phytomed 2015 Jul-Aug;5(4):288-97

Ahvaz Physiology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran. ; Department of Physiology, School of Medicine, Ahvaz Jundishapur University of medical Sciences, Ahavaz, Iran.

Objective: The aim of present study was to evaluate the effects of oral administration of gallic acid (GA) for 21 days alone and in combination with exercise on nerve conduction velocity and sensory and motor functions in rats with sciatic nerve crush.

Materials And Methods: Seventy adult male Wistar rats (250-300 g) were divided randomly into 7 groups with 10 in each: 1) Control (Cont), 2) Crushed + Vehicle (Cr +Veh), 3-5) Crushed + gallic acid (Cr+GA) (50, 100, and 200 mg/kg/2 mL, orally), 6) Crushed + exercise (Cr+Exe), and 7) Crushed + exercise + effective dose of gallic acid (Cr+Exe +GA200) for 21 days. In order to establish an animal model of sciatic nerve crush, equivalent to 7 kg of force pressed on 2-3 mm of sciatic nerve for 30 s, three times with 30 s intervals. Pain sense reflex in hot plate, motor coordination in rotarod, and sciatic nerve conduction velocity (SNCV) in all groups were tested. Data were analyzed using one-way ANOVA followed by Tukey's post hoc test and p<0.05 has assigned as the significant difference.

Results: Pain threshold was increased significantly in untreated crushed rats while motor function and SNCV were decreased in all groups with nerve crush (p<0.05, p<0.01, p<0.001 vs. control). Pain reflex latency was not changed in treated groups. Motor coordination and SNCV were improved in groups Cr+GA200 and Cr+Exe + GA200 (p<0.05, p<0.01 vs. Cr+Veh).

Conclusion: GA, dose-dependently, may have therapeutic potential to improve the peripheral nerve degeneration, which is most likely related, at least in part, to its antioxidant and therapeutic properties.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4587608PMC
October 2015

Roles of the nucleus accumbens (shell) in the acquisition and expression of morphine-induced conditioned behavior in freely moving rats.

Int J Prev Med 2014 Mar;5(3):262-8

Department of Physiology, Isfahan University of Medical Sciences, Isfahan, Iran.

Background: The nucleus accumbens (NAc) is a part of the rewarding cortico-mesolimbic dopamine (DA) pathway. This is a heterogeneous structure divided in two sub regions termed core and shell. DA function in the NAc is critical for goal-oriented behaviors, including those motivated by drug and brain stimulation reward. In the conditioned-place preference (CPP) paradigm, a test assessing animal's ability to associate drug-induced effects with environmental cause to quantify drug reward for example morphine.

Methods: In the present study, we investigated the influence of electrical stimulation with different current intensities on (25 and 100 µA) with and without an effective dose of morphine (0.5 and 5 mg/kg) on CPP.

Results: Subcutaneous administration of morphine 5 mg/kg produced significant CPP in comparison with saline group. Our findings also showed that electrical stimulation of NAc (100 µA) significantly (P < 0.01) suppressed morphine-induced CPP that reveals impaired learning and memory formation in the process of conditioning. We found that morphine-induced CPP can be successfully suppressed by current intensity (100 µA). It was probably due to decreasing of dopamine contents and its metabolites in the NAc. Current intensity (100 µA) in combination with ineffective dose of morphine (0.5 mg/kg) increased morphine-induced CPP probability via the prove reward system.

Conclusions: Since stimulation of dopaminergic neurons increases tendency to dependence to morphine, therefore in the present study, the stimulation of the NAc suppressed morphine-induced CPP that this shows impairment of learning and memory formation.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4018634PMC
March 2014

Effects of silver nano-particles on sperm parameters, number of Leydig cells and sex hormones in rats.

Iran J Reprod Med 2014 Feb;12(2):139-44

Department of Biology and Anatomy, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ; Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

Background: Nano-particles are extensively employed in most industries. Several studies have been started to explore the probable detrimental effects of nano-particles on human reproduction. However, there is insufficient and controversially evident of effects of silver nano-particles on sperm parameters and other reproductive indices.

Objective: Investigation of the effects of silver nano-particles on sperm parameters, sex hormones and Leydig cells in rat as an experimental model.

Materials And Methods: In this experimental study, 75 male prepubertal Wistar rats were categorized in five groups including control group and 4 experimental groups (n=15 in each group). The rats in the experimental groups were fed silver nano-particles (60 nm in dimension) with concentrations of 25, 50, 100, and 200 mg/kg/day. After 45 days (about one duration of spermatogenesis in rat), samples of blood were taken from the rats for testosterone, leuteinizing hormone (LH), and follicle stimulating hormone (FSH) assessments. Afterwards, the epididymis and the testis of each rat were dissected for analyzing sperm parameters and Leydig cells.

Results: The results demonstrated a statistically significant reduction in number of Leydig cells in experimental groups compared to control one. In addition, the data showed a reduction in testosterone and a rise in LH level which was more obvious in high doses (p<0.05); however, FSH level showed a reduction but it was not statistically significant. A significant decrease was also found in sperm motility and normal sperm morphology in the experimental groups compared to the control one.

Conclusion: Our results demonstrated that silver nano-particles, in addition to interruption in functions of sex hormones, can diminish the number of Leydig cells and sperm parameter indices. It should be noted that the effects of nano-particles on reproductive indices are dose-dependent.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4009568PMC
February 2014

Addition of Tempol in semen cryopreservation medium improves the post-thaw sperm function.

Syst Biol Reprod Med 2014 Aug 7;60(4):245-50. Epub 2014 Apr 7.

Department of Reproductive Biotechnology at Reproductive Biomedicine Research Center, Royan Institute for Biotechnology , ACECR, Isfahan , Iran .

Despite extensive research carried out for optimization and commercialization of sperm cryopreservation media, percentage of motility and viability remain low following cryopreservation. These adverse effects have been partially ascribed to reactive oxygen species (ROS) production during cryopreservation. Therefore, we proposed that addition of a cell permeable antioxidant like Tempol, with superoxide dismutase (SOD) mimetic action, may overcome these effects in an optimized commercially available cryo-protective medium. Therefore, semen samples were cryopreserved in the presence or absence of Tempol. A concentration of 5 μM Tempol was defined as optimal since it significantly improved motility and viability post thawing and reduced DNA fragmented sperm. In addition, percentage of ROS positive sperm was reduced. These effects of Tempol can be attributed to cell permeability characteristic and ability to reduce superoxide production both at intra- and extra-cellular levels. Tempol may hold the potential for clinical applications.
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http://dx.doi.org/10.3109/19396368.2014.897773DOI Listing
August 2014

N-acetylcysteine Prevents Kidney and Lung Disturbances in Renal Ischemia/Reperfusion Injury in Rat.

Int J Prev Med 2013 Oct;4(10):1139-46

Water & Electrolytes Research Center, Isfahan University of Medical Sciences, Isfahan, Iran ; Department of Biochemistry, Isfahan University of Payame-Noor, Isfahan, Iran.

Background: One of the most common causes of acute kidney injury (AKI) is kidney ischemia/reperfusion injury (IRI). The distant organ injury such as acute lung injury is one of the side effects of AKI or kidney IRI. In this study, we performed bilateral renal IRI in rats and the protective role of N-acetylcysteine (NAC) in kidney and lung was investigated.

Methods: Rats (n = 30) were randomly assigned to four experiment groups. The group 1 was assigned as sham-operated group. Before kidney IRI performance, the others groups were treated with saline (group 2), 150 mg/kg (group 3) or 500 mg/kg (group 4) of NAC, and the treatment were continued daily after IRI for next 3 days. At day 3, the all groups' animals were subjected for the measurements.

Results: The serum level of blood urea nitrogen (BUN) and creatinine (Cr) in the control group increased significantly (P < 0.05), and administration of NAC (150 mg/kg) decreased the serum levels of Cr and BUN. However, only the serum level of Cr decreased significantly (P < 0.05). NAC did not improve kidney weight and damage; however, its low dose (150 mg/kg) attenuated the lung injury score (P < 0.05) when compared with the control group. No significant differences were observed in lung water content and endothelial permeability, serum levels of malondialdehyde and nitrite between the groups.

Conclusions: Low dose of NAC as a protectant agent may protect the kidney function and lung tissue damage after kidney IRI.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3843300PMC
October 2013